L02 cells were purchased from ATCC, and cultured in DMEM (Gibco), supplemented with FBS 10% (Invitrogen, United States). Gecko library was used to infect 3 × 108 cells. The multiplicity of infection (MOI) was 0.1, and aimed to ensure that most cells received only 1 viral construct. The culture medium containing cells was supplemented with 10% FBS and 4 mM l- glutamic acid (Invitrogen), 10 μg/ml penicillin and streptomycin (Invitrogen, United States), followed by addition of the lentivirus to each dish with 8 μg/mL polybrene (Sigma). The cultures were incubated for 48 h, medium aspirated out, replaced with fresh DMEM supplemented with 1 μg/ml doxycycline, followed by incubation for 7 days. The cell population was created, with each target gene theoretically carrying a mutation of functional loss (Figure 1A , 2 ) (Wang et al., 2014b (link)).
L02 cells
Manufactured by American Type Culture Collection
Sourced in United States
L02 cells are a type of human liver cell line derived from normal adult liver tissue. They are commonly used in cell biology research and drug development as an in vitro model for studying liver function and toxicology.
Automatically generated - may contain errors
Lab products found in correlation
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
L glutamic acid, by Thermo Fisher Scientific (1 mentions)
Polybrene, by Merck Group (1 mentions)
213 ilb, by R&D Systems (1 mentions)
Thp 1, by American Type Culture Collection (1 mentions)
Lipopolysaccharides (lps), by Merck Group (1 mentions)
Hepg2 cell, by American Type Culture Collection (1 mentions)
Hk 2 cell, by American Type Culture Collection (1 mentions)
A549 cell, by American Type Culture Collection (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Puromycin, by neoFroxx (1 mentions)
Fulvestrant, by MedChemExpress (1 mentions)
7 protocols using l02 cells
1
Generation of Knockout Cell Line Library
2
Macrophage Polarization and Co-culture
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THP-1 (Human acute monocytic leukemia cell line) cells and L02 cells were purchased from ATCC. The cells were all cultured in RPMI 1640 containing 10% heat-inactivated FBS and 1% penicillin–streptomycin in a 5% CO2 atmosphere at 37 °C. THP-1 monocytes were differentiated into M0-like macrophages by incubation with 160 ng/ml phorbol 12-myristate 13-acetate (PMA, Sigma, P8139) for 48 h in RPMI medium. PMA-induced macrophages were polarized into M1 macrophages by incubation with 20 ng/ml of IFN-γ (R&D system, #285-IF) and 10 pg/ml of LPS (Sigma, #8630). Macrophage M2 polarization was obtained by incubation with 20 ng/ml of interleukin 4 (R&D system, #204-IL) and interleukin 13 (R&D system, #213-ILB). In the co-culture experiments, 0.5 × 106 THP-1 monocytes were cultivated and differentiated for 48 h before being incubated respectively with 0.5 × 106 L02-SCR or L02-Sh cells in 6-well Transwell plates for 48 h37 (link) .
3
Oxidative Stress Modulates Hepatic PGC-1α and IL-6
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human non-tumor hepatic L-02 cells were obtained from ATCC (Manassas, VA, USA). Mouse primary hepatocytes were isolated from C57BL/6 mice as described previously [19 (link), 40 (link)]. L-02 cells and primary hepatocytes were cultured in Dulbecco’s modified Eagle’s medium supplemented with 10% of fetal bovine serum (FBS) at 37 °C in a humidified incubator of 5% CO2. The cells were transfected with a PGC-1α (accession code: NM-013261) CRISPR activation plasmid, PGC-1α siRNAs, the control plasmid or siRNA using Lipofectamine 2000, following the manufacturer’s protocols. After transfection for 24 h, the cells were treated with 300 or 100 μM H2O2 for 48 h and harvested for analysis of cellular PGC-1α and IL-6 expression.
4
Cell Culture Protocols for Assays
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HK-2 cells (Human Kidney Tubular Epithelial Cells), L02 cells (Hepatocytes), ES-2 and OVCAR8 cells (Human Ovarian Carcinoma Cells), HepG2 cells (Hepatocellular Carcinoma Cells), and A549 cells (Lung Carcinoma Cells) were purchased from American Type Culture Collection (ATCC, Manassas, VA). HK-2 and OVCAR8 cells grown in 1640 medium (10% FBS and 1% penicillin/streptomycin), ES-2 cells grown in McCoy’s 5A medium, L02 and HepG2 cells grown in DMEM medium, A549 cells grown in F-12 medium were at 37 °C under a humidified atmosphere with 5% CO2.
5
Cultivation of Hepatocellular Carcinoma Cell Lines
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Human hepatoma HepG2 and HepG2215 cell lines and normal human liver L02 cells were purchased from American Type Culture Collection (ATCC; Manassas, VA). HCC HuH-7 cells were purchased from Procell Life Science & Technology Co., Ltd (Procell; Wuhan, China). HepG2215 cells are HBV-transfected HepG2 cells that can replicate HBV [27 (link)]. The cells were cultured in Dulbecco's Modified Eagle Medium (DMEM) (Gibco, USA) containing 10% fetal bovine serum (Gibco, USA). All cells were cultured in a carbon dioxide incubator having a humid atmosphere of 37°C and 5% CO2.
6
Estrogen-Modulated Liver Cell Culture
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L02 cells (normal human liver cells) were purchased from the American Type Culture Collection (ATCC). L02 cells were cultured in high-glucose DMEM, phenol red free DMEM (PRF DMEM) medium (HyClone) containing 10% fetal bovine serum (FBS) or estrogen charcoal-stripped FBS (CS FBS), and 1% penicillin/streptomycin (100 μg ml−1) at a constant temperature (37 °C) in a humidified incubator (5% CO2). To ensure that the experimental results were not influenced by exogenous estrogen levels, PRF DMEM medium and CS FBS were used in our experiments that were conducted in preparation of CS FBS and was followed based on a prior work (de Faria et al. 2016 (link); Simoncini et al. 2005 (link)). Fulvestrant (ICI 182780, MedChemExpress, 100 nM) was used for estrogen receptor inhibition. L02 cells were exposed to lentivirus (LV)-HNF1b (Genechem Co., Ltd., Shanghai) or LV-shPPARγ (TianYuCheng Co., Ltd., Xi’an) for 12 h and then selected with puromycin (BioFroxx, Germany) for 12 h to establish a cell line with stable overexpression of HNF1b. L02 cells were treated with 1 or 10 μM BPA for 48 h.
7
Cultivation of Normal Human Hepatic L02 Cells
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Normal human hepatic cell line (L02 cells) was purchased from the American Type Culture Collection (ATCC, Manassas, VA), and maintained in RPMI 1640 medium (containing 10 % (v/v) fetal bovine serum (FBS), penicillin of 100 U/mL, and streptomycin of 100 g/mL).
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